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  • Title: Effects of methanol on embryonic mouse palate in serum-free organ culture.
    Author: Abbott BD, Logsdon TR, Wilke TS.
    Journal: Teratology; 1994 Feb; 49(2):122-34. PubMed ID: 8016743.
    Abstract:
    Methanol has widespread applications in industry and manufacturing and is under consideration as an alternative automotive fuel. Human exposure to methanol would be expected to increase if applications expand in coming years. Methanol has been shown to be a reproductive and developmental toxicant in the rodent, producing cleft palate in the CD-1 mouse. Developmental toxicity has also been demonstrated in vitro for rat and mouse embryos in whole embryo culture. The present study examines the developmental toxicity of methanol in the palate using a serum-free organ culture model. Gestation day 12 CD-1 mouse embryos were dissected and mid-craniofacial tissues were cultured in BGJ medium at 37 degrees C for 4 days with medium changes at 24 hr intervals. Cultures were exposed to methanol from 0-20 mg/ml for 6 hr, 12 hr, 1 or 4 days. Some cultures were exposed to ethanol for 4 days at doses ranging from 0-15 mg/ml. All cultures were gassed with a 50% O2, 5% CO2, and 45% N2 upon addition of fresh medium and prior to the addition of alcohol. Following organ culture the craniofacial explants were examined for effects on morphology, fusion, proliferation, and growth. Incidence and completeness of palatal fusion decreased with increasing exposure. Depending on the concentration and duration of methanol exposure, the medial epithelium either degenerated completely or remained intact in unfused palates and either condition would interfere with fusion. Cellular proliferation appeared to be a specific and sensitive target for methanol as craniofacial tissues responded to methanol with reduction in total DNA content at an exposure that did not affect total protein. However, both DNA and protein decreased with increasing exposure to methanol. Incorporation of thymidine decreased significantly after 4 day exposure and autoradiography of 3H-thymidine (TdR) demonstrated exposure-dependent reduction in proliferation of palatal mesenchymal cells. Ethanol decreased fusion score, total protein, and DNA, but 3H-TdR/DNA was not significantly changed. In general the ethanol was more potent than methanol for inhibition of protein and DNA synthesis and palatal fusion. This study demonstrated that methanol can selectively affect specific sensitive cell populations and has effects on proliferation and cell fate.
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